Total Phenolic Content, Antioxidant, Cytotoxicity and Hepatoprotective Activities of Aqueous Extract of Channa striatus (Haruan)

Abstract

Antioxidant has been the important approaches to reduce the development of cancer disease and play a role as a protective against liver toxicity. Channastriatus (haruan) have been used traditionally oral remedy for wound healing among women after child birth. However, there is little scientific evidence or research yet regarding the cytotoxicity and hepatoprotective activity of C.striatus. Thus, the aims of this study are to determine potential total phenolic content (TPC), antioxidant, cytotoxicity and hepatoprotective effect of C.striatus (Haruan) extract. For this present study, aqueous and lipid extract of was prepared using chloroform and methanol solvent in a ratio of 2:1. Folin-Ciocalteu was used to quantify TPC and three antioxidant assays were used to determine antioxidant activity, including 2,2-diphenyl-picrylhydrazyl (DPPH) assay, azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) and ferric reducing ability of power (FRAP). For cell viability assay, HepG2 cell lines were seeded in 96-well plates and were treated with various concentration of aqueous extract of C.striatus, AECS (0, 0.00001, 0.0001, 0.001, 0.01, 0.1, 1, and 10 mg/ml) at 24, 48 and 72 hours. MTT assay has been used to measure HepG2 viability by using ELISA microplate reader. For hepatoprotective study, thirty six of adult male Sprague-Dawley rats will be divided into six groups of six rats each (n=6): G1:control (10%DMSO), G2:negative control (10% DMSO), G3:positive control (silymarin-100 mg/kg),G4: C.striatus (50mg/kg), G5:C.striatus (150mg/kg) and G6:C.striatus (450 mg/kg). The extract was given orally for 1 week. Acetaminophen,AAP (3g/kg) was induced orally from group 2 to 6 after 7 days of treatment. Blood collection was analyzed for liver function test and then the rats were sacrificed for histopathologically study. In TPC assay, AECS was observed to have higher content of phenolic (12799.33±237.90) compared to lipid extract of C.striatus, LECS (515.33±160.75). This indicates that, AECS has higher scavenging activity in DPPH and ABTS assay with EC 50 (64.93±10.78) and (4687±0.67)respectively in comparison to LECS with EC 50 (0.1513±0.046) and (93333.33±11.25) respectively. Additionally, AECS also consists of reducing potency since it has higher ability to reduce ferric ion compared to lipid extract of Channa striatus.For MTT assay, a significant decrease the percentage of HepG2 viability in a dose dependent manner was observed after HepG2 treated with various concentration of AECS at 24, 48 and 72 hours. The result showed no IC 50 value of HepG2 obtained at 24 hours while IC 50 value of AECS were 0.85 ± 0.26 and 0.1 ± 0.04mg/ml at 48 and 72 hours respectively. In vivo study, all groups pretreated of rats with AECS has shown significant decrease (p <0.05) in the level of ALT, AST, AST and histological scoring of liver when comparing with acetaminophen group. In conclusion, AECS has higher TPC and antioxidant activity than LECS. Besides that, AECS exhibited potential cytotoxicity effect towards HepG2 cell lines in both dose and time dependent manner and also hepatoprotective effect at lowest dose of AECS (50 mg/kg). Further studies are required for fully elucidation on the mechanism of cytotoxicity and hepatoprotective activity of AECS.